Dioecious <Emphasis Type="Italic">Clusia nemorosa</Emphasis> achieves pollination by combining specialized and generalized floral rewards

Plants often combine multiple strategies to achieve pollen transfer. The dioecious Clusia nemorosa (Clusiaceae) produces floral resin, a specialized reward for resin-collecting bees. It also exhibits floral automimicry with female flowers mimicking male flowers in order to attract pollen-collecting insects. We observed the hourly visitation frequency and behaviour of bee visitors on two rock savanna sites in French Guiana. The major strategy in pollen transfer and the variation in visitation rates among visitor species were examined. We hypothesized that the visitation rate will vary between the floral sexes and degree of this variation will differ between the two-reward systems. We found no evidence for visits being exclusively related to resin collection, which we expected to be the principal strategy in pollen transfer. Deceit pollination appeared to have minor importance and seemed to be locally associated with the site, where demand for pollen was greater. Flower visits that probably facilitated most pollen transfer, combined resin and pollen. The pollinators involved in this system collected pollen from male flowers and resin from female flowers. The fruit set was not particularly low (44.19%) so offering different rewards by different flower genders has not constrained reproductive success in this system. Pronounced variation in visitation rates between sexes was not related to the pollination mechanisms but to the demand for the two rewards. Overall demand for pollen was greater than demand for resin. Female flowers were visited much less frequently than male flowers. Peak pollen collection occurred in the morning, i.e., as soon as the resource was available, while resin was collected throughout the day. There was a local variability in demand for the two resources because both the visitation rate and the behaviour of foraging bees varied between the two observation sites.     

The behaviour of male orchid bees (Apidae,Hymenoptera, Insecta) and the question of leks

The male territories of two species of orchid bees, Eulaema meriana and Euglossa imperialis, are described. These territories consist of a perch, where the males display on the trunk of a tree, and a route flown from and back to the perch. Territories are located in treefalls or other large light gaps in tropical forest. The territories of these two species differ in the height of the perch and amount of light reaching it, size of the perch tree, and period of activity. A definition of lek is given. These male euglossines form facultative leks in large treefalls. The relationships between male territories, female foraging ranges, and collection of aromatic material by males are discussed.     

Juggling with volatiles: exposure of perfumes by displaying male orchid bees

Male orchid bees (Euglossini) pollinate 10% of the neotropical orchid flora while collecting floral scents, which they store and accumulate in hind tibial pouches. The purpose of these fragrances is unclear, as is the context, timing and mechanism of their possible exposure. Here we show for the first time that males expose and relocate their fragrances during courtship display. We present high-speed video analyses revealing an intricate and repetitive leg movement performed by displaying male Euglossa cognata. The behavior involves several morphological structures of hitherto unknown function and suggests transfer of substances from the hind tibia to a contralateral mid-tibial tuft of hairs. Body-side-specific fluorescent dye application and consecutive detection of signals on males after display confirmed this transfer. Deposited on the mid-tibial tufts, the fragrances are ideally placed in order to become ventilated by jugal combs on the wing bases, as previously suggested by Bembé (in Apidologie 35:288–291, 2004). Being clearly distinct from motor patterns involved in fragrance collection, the described movement is continuously performed by displaying males, suggesting an equally continuous exposure of volatiles. Although the findings strengthen, the view that the volatiles serve as attractants in the context of mating behavior, the signal addressee, conspecific males or females, has yet to be found.Electronic Supplementary Material Supplementary material is available for this article at     

Molecular complexity from aromatics: synthesis and photoreaction of endo-tricyclo[5.2.2.0(2,6)]undecane--a stereoselective route to tricyclic framework of protoilludanes

A stereoselective route towards protoilludanoids from simple aromatic precursor is described. The methodology involves in situ generation of spiroepoxycyclohexadienone and cycloaddition with cyclopentadiene and photochemical 1,3-acyl shift.     

Ring-cleaving dioxygenases with a cupin fold

Ring-cleaving dioxygenases catalyze key reactions in the aerobic microbial degradation of aromatic compounds. Many pathways converge to catecholic intermediates, which are subject to ortho or meta cleavage by intradiol or extradiol dioxygenases, respectively. However, a number of degradation pathways proceed via noncatecholic hydroxy-substituted aromatic carboxylic acids like gentisate, salicylate, 1-hydroxy-2-naphthoate, or aminohydroxybenzoates. The ring-cleaving dioxygenases active toward these compounds belong to the cupin superfamily, which is characterized by a six-stranded β-barrel fold and conserved amino acid motifs that provide the 3His or 2- or 3His-1Glu ligand environment of a divalent metal ion. Most cupin-type ring cleavage dioxygenases use an Fe(II) center for catalysis, and the proposed mechanism is very similar to that of the canonical (type I) extradiol dioxygenases. The metal ion is presumed to act as an electron conduit for single electron transfer from the metal-bound substrate anion to O(2), resulting in activation of both substrates to radical species. The family of cupin-type dioxygenases also involves quercetinase (flavonol 2,4-dioxygenase), which opens up two C-C bonds of the heterocyclic ring of quercetin, a wide-spread plant flavonol. Remarkably, bacterial quercetinases are capable of using different divalent metal ions for catalysis, suggesting that the redox properties of the metal are relatively unimportant for the catalytic reaction. The major role of the active-site metal ion could be to correctly position the substrate and to stabilize transition states and intermediates rather than to mediate electron transfer. The tentative hypothesis that quercetinase catalysis involves direct electron transfer from metal-bound flavonolate to O(2) is supported by model chemistry.     

Identification of the Proton Source for the Microbial Reductive Dechlorination of 2,3,4,5,6-Pentachlorobiphenyl

Microbially mediated reductive dechlorination of polychlorinated biphenyls (PCBs) in anaerobic sediments has been observed during laboratory experiments. Reductive dechlorination is a two-electron transfer reaction which involves the release of chlorine as a chloride ion and its replacement on the aromatic ring by hydrogen. The exact mechanism of the electron transfer for PCBs is unknown; however, this work shows that the source of the hydrogen atom is the proton (H⁺) from water.     

Linnaeus's butterflies (Lepidoptera: Papilionoidea and Hesperioidea)

The names of all the species of butterflies described by Linnaeus under "Papilio" are researched. Of the 305 names treated, 243 (c. 80%) are currently valid as specific (241) or subspecific (2), 29 are junior synonyms, 14 are invalid (one of these applying to a fake), and for 13 the identity is unknown or uncertain. Six species of moths misidentified by Linnaeus as butterflies are cited in the study, but details are not included. One hundred and fifty-two lectotypes have been designated, representing about 56% of the sum of the valid names and junior synonyms. Of these, 99 were selected from specimens in the Linnean Society of London, 52 from Queen Ludovica Ulrica's collection, Uppsala, and one lectotype is a Petiver specimen from the collection of Sir Hans Sloane. Linnaeus described at least five species, possibly eight, from the literature alone. All Linnaean material examined is documented, as are 'subsequent' specimens that are associated with Linnaean material. Synonymy and homonymy are discussed and presented, as are the identities of type localities. Care has been taken to achieve a practical balance between Linnaean and current species identities. Linnaean material studied included specimens from The Linnean Society of London, Museum Ludovicae Ulricae (Uppsala University), the Clerck and De Geer collections in the Naturhistoriska Riksmuseet, Stockholm, and the collection of James Petiver, now part of the collection of Sir Hans Sloane housed in The Natural History Museum, London.     

Histidine and Aromatic Permeases of Salmonella typhimurim

Mutants defective either in the histidine permease (hisP) or in the aromatic permease (aroP) were isolated in Salmonella typhimurium and were characterized. The hisP locus had a 49% linkage to purF by phage transduction. The aroP locus was close to proA. Merozygotes diploid for the hisP gene were constructed by episomal transfer, and hisP(+) was dominant over hisP. The properties of merozygotes are described and discussed. A method for the selection of revertants of hisP mutants was devised. By this method, one of the hisP mutants was characterized as an amber mutant. The specificity of the aromatic permease was investigated by using as substrates analogues of the aromatic amino acids and of histidine.     

Preparation of immobilized monomeric actin and its use in the isolation of protease-free and ribonuclease-free pancreatic deoxyribonuclease I

A procedure is described for the immobilization of monomeric actin so that about 30% of the immobilized protein is competent to bind the monomeric-actin-binding proteins bovine pancreatic deoxyribonuclease I and chicken villin. The intact tertiary structure of the immobilized actin is required to bind these proteins. Using this resin, a method has been developed for the affinity purification of pancreatic deoxyribonuclease I on a reusable actin column. It involves the binding of deoxyribonuclease I to immobilized actin, extensive washing of the column, followed by elution of the bound deoxyribonuclease I with 10 M formamide. After removal of the formamide, the deoxyribonuclease I has a higher specific activity than the starting material and contained no detectable protease or ribonuclease contamination. This preparation should find considerable application in molecular genetic studies where the enzyme is needed free of these particular contaminants. The affinity column should also be useful for the isolation of other, physiologically relevant, monomeric-actin-binding proteins.     

Liquid phase synthesis of arylamines and its application to the benzimidazolone via nucleophilic aryl substitution.

A method for soluble, inexpensive polymer-supported synthesis of aryl amines and benzimidazolone on the basis of nucleophilic aryl substitution (S(N)Ar) is described. This method involves a direct coupling reaction between resin bound aryl fluoride and amines at ambient temperature. The products are isolated in quantitative yields and excellent purity by simple precipitation and washing. This liquid phase method proves to be a useful tool for constructing combinatorial arylamine and benzimidazolone libraries.     

The Linnaean Species of the Families Fanniidae, Anthomyiidae and Muscidae (Insecta: Diptera)

Linnaeus described eleven species in the genera Musca and Conops that are now placed in the Fanniidae, Anthomyiidae and Muscidae. A study is presented of the types and other material of these three families in his collection.     

The laparoscope in follicular oocyte collection and gamete intrafallopian transfer and fertilization (GIFT)

Laparoscopic recovery of bovine follicular oocytes was studied. The collection of oocytes from the superovulated bovine ovary was maximized by standardizing the collection technique. The technique was highly successful, with a 79% oocyte recovery rate of the follicles aspirated. Collected oocytes were transferred to the inseminated recipient's oviduct with a minimum of trauma through the laparoscope. This gamete intrafallopian transfer and fertilization (GIFT) resulted in multiple embryo recovery in the cow. Oviductal catheterization and the potential of GIFT are described and discussed.     

Rapid assessment of rivers using macroinvertebrates: A procedure based on habitat-specific sampling,family level identification and a biotic index

Abstract This paper describes a simple and inexpensive procedure for the rapid biological assessment of water quality in rivers and streams in eastern Australia. The procedure involves the standardized collection of samples of 100 macroinvertebrates from defined habitat types within a water body. Specimens are identified to family level only and a biotic index is calculated. Proposed future testing and evaluation are described, and the limitations of the rapid approach are discussed.     

Electrophoretic transfer of proteins from fixed and stained gels

A procedure by which sodium dodecyl sulfate gels can be fixed and stained with Coomassie blue and subsequently transferred to nitrocellulose for immunostaining is outlined. The procedure involves the complete removal of the stain followed by equilibration of the gel in sodium dodecyl sulfate running buffer. The efficiency of transfer is comparable to unfixed gels and the protein pattern of the transfer appears to be sharper, presumably due to less diffusion during the transfer process. The procedure does not affect the antigenicity of the proteins that have been examined by subsequent immunostaining. This method is particularly useful for situations in which sample size or concentration are limiting factors resulting in insufficient material for duplicate gels.     

Ovarian and uterine lymphatic drainage in Australian flying-foxes (genus Pteropus,suborder Megachiroptera)

Ovarian lymphatics of flying-foxes were traced to determine if they could transport hormones directly from ovary to ipsilateral uterine horn, thereby stimulating the localised endometrial growth which is characteristic of these animals. Intra-ovarian injections of ink and serial histological sections did not reveal any such connection. All major ovarian lymphatics and those from the cranial tip of each uterine horn drain cranially, terminating in 1 or 2 lymph nodes lying caudal to the ipsilateral kidney. For much of their course, the major ovarian lymphatics run in the adventitia of the ovarian venous sinus. This sinus encloses the coiled ovarian artery, which provides the major blood supply to the cranial end of the ipsilateral uterine horn. Some fine ovarian lymphatics run in the adventitia of the coiled ovarian artery. The enclosure of the coiled ovarian artery by the ovarian venous drainage is thought to provide the main route for transfer of steroids from ovarian vein to ovarian artery and thence to ipsilateral uterine horn. The ovarian lymphatics described here do not bypass the vascular pathway but provide an additional route for counter-or cross-current transfer of ovarian steroids to the ovarian arterial supply to the uterus.     

Preparation of serial sections of arthropods using 2,2-dimethoxypropane dehydration and epoxy resin embedding under vacuum

Improved methods are described for anatomical investigation of small insects and other arthropods using serial semithin sections. The specimens were dehydrated with acidified 2,2-dimethoxypropane and embedded in ERL 4206 epoxy resin under vacuum. This procedure ensures good resin impregnation of thin, long body compartments and appendages. Furthermore, it produces excellent overall preservation of the specimen and its fragile anatomical structures. This procedure saves time and gives excellent results when sectioning difficult arthropod material. A continuous recording of serial semithin sections is possible when diamond knives are used.     

Batch-To-Batch Variation of Chelex-100 Confounds Metal-Catalysed Oxidation. Leaching of Inhibitory Compounds from A Batch of Chelex-100 and Their Removal by a Pre-Washing Procedure

Removal of adventitious redox-active metals from buffers by treatment with Chelex resin is a widely used procedure in free radical research. Use of a new batch of Chelex-100 resin in our laboratory coincided with a sudden inability to oxidise low-density lipoprotein with copper. We found that copper-mediated oxidation of ascorbate in water treated with the same batch of Chelex was inhibited when compared with untreated water and water treated with a different batch of the resin. Washing the Chelex removed the inhibitory effect suggesting that material was leaching from the resin. The washing procedure for Chelex-100 described is simple and can be scaled up. Oxidation of ascorbate with low concentrations of copper can be used to test the quality of batches of the resin.     

Preparation of serial sections of arthropods using 2,2-dimethoxypropane dehydration and epoxy resin embedding under vacuum.

Improved methods are described for anatomical investigation of small insects and other arthropods using serial semithin sections. The specimens were dehydrated with acidified 2,2-dimethoxypropane and embedded in ERL 4206 epoxy resin under vacuum. This procedure ensures good resin impregnation of thin, long body compartments and appendages. Furthermore, it produces excellent overall preservation of the specimen and its fragile anatomical structures. This procedure saves time and gives excellent results when sectioning difficult arthropod material. A continuous recording of serial semithin sections is possible when diamond knives are used.     

Synthesis of bis-armed amino acid derivatives via the alkylation of ethyl isocyanoacetate and the Suzuki–Miyaura cross-coupling reaction

Summary. Two synthetic routes to bis-armed-α-amino acid derivatives are described. The first route involves alkylation of dibromo derivatives with ethyl isocyanoacetate under phase-transfer catalysis (PTC) conditions. The second route uses a palladium-mediated Suzuki–Miyaura cross-coupling reaction between a DL-4-boronophenylalanine derivative and aromatic diiodo (or dibromo) compounds.     

A NEW SPECIES OF PICEA BASED ON SILICIFIED SEED CONES FROM THE OLIGOCENE OF WASHINGTON

Picea eichhornii n. sp. is described from anatomically preserved seed cones. The fossils are from the Early Oligocene Jansen Creek Member of the Makah Fm. which is exposed along the northern shore of the Olympic Peninsula, Washington. The cones are at least 5.5 cm long and up to 3.5 cm in diameter. The cone axis is 4–6 mm in diameter and contains a pith made up of thick-walled parenchyma cells. Resin canals occur in a single ring in the secondary xylem in some specimens but are absent in others. The cortex is mostly parenchymatous and contains numerous large axial resin canals that branch to supply the bract and scale. Vascular traces to each scale and its subtending bract diverge separately from the vascular cylinder of the cone axis. The bract is tongue-shaped and keeled at its base. It is 5 mm wide and up to 9 mm long. The bract trace fades out before entering the bract base while two resin canals extend into the bract base. The ovuliferous scale is about 2.3 cm long and has a thin, probably papery, apex. Resin canals of the scale occur abaxial to the vascular tissue in the scale base, but some bend around the margins of the vascular strand to become adaxial outward. About 20 resin canals occur in the abaxial scale sclerenchyma, and this is the main anatomical feature that distinguishes these cones as a new species. There are less than 14 such canals in cones in a reference collection of 15 modern species and in the two fossil species known from anatomically preserved material. While the new species adds to our knowledge of the diversity of Cenozoic Picea, its affinities within the genus remain undetermined.